CN109207388B - Preparation and application of glass fiber/high polymer composite immobilized microbial filler - Google Patents

Preparation and application of glass fiber/high polymer composite immobilized microbial filler Download PDF

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CN109207388B
CN109207388B CN201710531167.4A CN201710531167A CN109207388B CN 109207388 B CN109207388 B CN 109207388B CN 201710531167 A CN201710531167 A CN 201710531167A CN 109207388 B CN109207388 B CN 109207388B
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photosynthetic bacteria
glass fiber
filler
fiber balls
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CN109207388A (en
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张国亮
张旭
秦磊
孟琴
刘秋花
范铮
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Zhejiang University of Technology ZJUT
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    • C12N1/00Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00Biological treatment of water, waste water, or sewage
    • C02F3/34Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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    • C12N11/00Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
    • C12N11/02Enzymes or microbial cells immobilised on or in an organic carrier
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    • C12N11/00Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
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    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2209/00Controlling or monitoring parameters in water treatment
    • C02F2209/08Chemical Oxygen Demand [COD]; Biological Oxygen Demand [BOD]

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Abstract

The invention discloses a preparation method of a glass fiber/high polymer composite immobilized microbial filler, belonging to the field of water treatment. The technical scheme of the invention is to provide a capsule type immobilized filler which is easy for the growth of microorganisms and has the characteristics of the immobilized filler, and the specific method comprises the following steps: (1) obtaining a high-efficiency photosynthetic bacteria strain through enrichment culture; (2) mechanical hot ligation of glass fiber balls; (3) photosynthetic bacteria are adsorbed and grown in the glass fiber balls; (4) preparing precursor solution of high polymer coating materials (polyvinylidene fluoride, polysulfone, polyolefins and the like); (5) and (3) synthesis of the photosynthetic bacteria immobilized filler. The filler prepared by the microorganism immobilization method has stronger mechanical property, and overcomes the problems of poor mass transfer performance and low light utilization rate of the traditional embedding filler due to the porosity and light transmittance of the glass fiber ball.

Description

Preparation and application of glass fiber/high polymer composite immobilized microbial filler
Technical Field
The invention belongs to the technical field of water treatment, and particularly relates to a preparation method of an efficient fixed photosynthetic bacteria filler.
Background
Photosynthetic bacteria (PSB) are a type of microorganism that uses light as an energy source and can use organic matter, sulfide, ammonia, etc. in nature as a hydrogen donor and carbon source to perform photosynthesis under anaerobic illumination or aerobic dark conditions. Because photosynthetic bacteria have small thalli and are difficult to naturally settle, two problems of thallus loss and solid-liquid separation exist in practical application. In order to solve the two problems, fresh thalli need to be continuously cultured and added, and solid-liquid separation treatment work is also needed, so that the treatment process flow is complicated, the treatment cost is increased, and the popularization and the application of the microbial biomass in production are seriously influenced. The traditional immobilization method has a plurality of applications, but filler prepared by polyvinyl alcohol, sodium alginate, agar, gelatin and the like as embedding agents has poor mechanical properties, embedding materials can influence the diffusivity of substrates and products, most of the materials have poor mechanical properties and are easy to break, so that the application of the method is limited, and the adsorption method mostly uses porous materials as carriers to attach bacteria thereon, so that the bacteria are easy to fall off from the carriers under the action of water power. Therefore, the invention uses inorganic carrier material glass fiber ball as carrier, polyvinylidene fluoride or polysulfone as porous shell, the prepared immobilized filler not only has better mass transfer property, but also has high mechanical strength and is not easy to break.
Disclosure of Invention
The invention aims to solve the main problems in the prior bacterial immobilization, and provides an immobilized microorganism technology, wherein photosynthetic bacteria are adsorbed in a glass fiber ball by an adsorption method, and a layer of film is coated on the outer surface of the glass fiber ball by a coating method, so that an immobilized filler is prepared.
The purpose of the invention is realized by the following technical scheme:
a composite type photosynthetic bacteria immobilized filler is prepared by the following method:
(1) culturing photosynthetic bacteria in a photosynthetic bacteria liquid culture medium under the condition of illumination and anaerobic conditions until the concentration is 1-2g/L in terms of dry cell mass to obtain a photosynthetic bacteria enrichment culture solution, adding glass fiber balls, and adsorbing the photosynthetic bacteria in pore channel structures of the glass fiber balls through the adsorption action of the glass fiber balls to obtain the photosynthetic bacteria adsorption glass fiber balls;
(2) adding the high polymer into an organic solvent, stirring and dissolving at the temperature of 20-40 ℃ to obtain a coating reagent; the high polymer is polysulfone or polyolefin; the mass fraction of the high polymer in the coating reagent is 8-20%;
(3) and (2) immersing the glass fiber balls adsorbing the photosynthetic bacteria obtained in the step (1) in the coating reagent prepared in the step (2), stirring, shaking uniformly, taking out, placing in deionized water, standing to form a film-shaped porous coating layer on the surfaces of the glass fiber balls, taking out, and washing with distilled water to obtain the composite photosynthetic bacteria immobilized filler.
Further, the final concentration of the photosynthetic bacteria liquid culture medium of the invention comprises the following components: 0.5g/L yeast extract and 0.5g/L, CH peptone3COONa 3g/L、NH4Cl 0.1g/L、NaCl 0.5g/L、NaHCO3 0.5g/L、K2HPO4 0.2g/L、MgSO4·7H2O 0.1g/L、CaCl20.1g/L, water as solvent, and initial pH of 7-9.
Further, the preparation method of the glass fiber ball comprises the following steps: sterilizing glass fibers, performing mechanical hot ligation on the glass fibers to prepare spheres with uniform sizes, and cleaning and drying the spheres to obtain the glass fiber spheres.
Further, in the step (2) of the present invention, the organic solvent is N, N-dimethylformamide, N-dimethylacetamide, or N-methylpyrrolidone.
Further, in step (2) of the present invention, the polymer is preferably polyetherimide, polysulfone, polyethersulfone, polyacrylonitrile, polytetrafluoroethylene, or polyvinylidene fluoride.
Still further, the high polymer in the step (2) of the present invention is more preferably polysulfone or polyvinylidene fluoride.
Further, the volume of the photosynthetic bacteria enrichment culture solution in the step (1) is 100-1000 mL/g based on the mass of the added glass fiber balls.
Further, the solid-liquid separation in the step (1) is realized by centrifugation at the rotating speed of 10000-12000 rpm.
Further, the photosynthetic bacteria of the present invention are Rhodopseudomonas palustris photosynthetic bacteria.
Further, the invention provides an application of the composite photosynthetic bacteria immobilized filler in degrading high-salinity wastewater COD.
The invention has the beneficial effects that: the filler prepared by the microorganism immobilization method has stronger mechanical property, and overcomes the problems of poor mass transfer performance and low light utilization rate of the traditional embedding filler due to the porosity and light transmittance of the glass fiber ball.
Drawings
FIG. 1: example 1 an immobilized filler prepared from polyvinylidene fluoride (PVDF) and Polysulfone (PSF) of example 3;
FIG. 2: the degradation effect of the immobilized filler on COD.
FIG. 3: example 3 and comparative example 2 effect on COD degradation.
Detailed Description
The invention will be further described with reference to specific examples, but the scope of the invention is not limited thereto.
The photosynthetic bacteria of the present invention are preferably Rhodopseudomonas (Rhodopseudomonas palustris) photosynthetic bacteria, and the photosynthetic bacteria seed solution is purchased from Dinglong group of Zhejiang.
The sterile water adopted by the invention is deionized water treated by a high-pressure steam sterilization pot, and the treatment condition is that the temperature is 121 ℃ and the pressure is maintained for 30 minutes under the condition of 0.105 Mpa.
Example 1:
preparation of glass fiber balls
Sterilizing 3g of glass fiber in a high-pressure steam sterilization pot, then performing mechanical heat ligation on glass fiber balls with different filling densities to prepare glass fiber balls with uniform sizes, cleaning in sterile water, and drying to obtain the glass fiber balls.
Second, culture of photosynthetic bacteria
1. Configuring photosynthetic bacteria liquid culture medium
Weighing yeast extract 0.5g, peptone 0.5g, and CH3COONa 3g、NH4Cl 0.1g、NaCl 0.5g、NaHCO30.5g、K2HPO4 0.2g、MgSO4·7H2O 0.1g、CaCl2Adding 0.1g of the mixture into 1000mL of distilled water, adjusting the pH value to 7 by using 10% of Na0H and 10% of HCI in percentage by mass, and uniformly stirring to obtain the photosynthetic bacteria liquid culture medium.
2. Enrichment culture of photosynthetic bacteria
200ml of photosynthetic bacteria liquid culture medium is prepared, the photosynthetic bacteria liquid culture medium is poured into a 250 ml conical flask for high-pressure steam sterilization, wherein the sterilization temperature is 115 ℃, the sterilization time is 30 minutes, and the conical flask is taken out and cooled to the room temperature. Transferring photosynthetic bacteria stored in the 7-ring solid culture medium into a conical flask by using an inoculating ring, uniformly mixing, wrapping the periphery of the conical flask by using gauze, placing the conical flask into an incubator, and performing illumination anaerobic culture for 48 hours under the condition that the light intensity is 2000lux, wherein the culture temperature is 30 ℃, so as to obtain the photosynthetic bacteria enrichment culture solution.
Preparation of photosynthetic bacteria immobilized filler
1. Preparation of coating solution
And adding polyvinylidene fluoride into the N, N-dimethylformamide solution, stirring and dissolving to obtain the polyvinylidene fluoride solution.
2. Adsorption of glass fiber balls to bacteria
0.5g of glass fiber balls are weighed, washed clean, dried and added to 100mL of photosynthetic bacteria with the concentration of 1g/L for adsorption.
3. Preparation of immobilized fillers
And (3) placing the glass fiber balls adsorbed with 0.3g of bacteria in the prepared polyvinylidene fluoride solution, stirring, shaking uniformly, taking out, placing in deionized water, standing for 4min, taking out, and washing with distilled water.
Fourthly, simulating high-salinity wastewater indexes: salinity of 5%, COD: 2900 mg/L.
Fifth, activity detection of immobilized filler
0.5g of immobilized filler is put into 200mL of high-salinity wastewater and treated for 72h under the conditions that the temperature is 30 ℃ and the illumination intensity is 2000 lux.
Two sets of experiments were set up, the difference being that the concentration of the polymer in the coating solution was 8% and 20%, respectively, and it can be seen from fig. 2 that the degradation of the high salinity wastewater COD by the immobilized photosynthetic bacteria was 88% and 82%, respectively.
Example 2:
preparation of glass fiber balls
Sterilizing 3g of glass fiber in a high-pressure steam sterilization pot, then performing mechanical heat ligation on glass fiber balls with different filling densities to prepare glass fiber balls with uniform sizes, cleaning in sterile water, and drying to obtain the glass fiber balls.
Second, culture of photosynthetic bacteria
1. Configuring photosynthetic bacteria liquid culture medium
Weighing yeast extract 0.5g, peptone 0.5g, and CH3COONa 3g、NH4Cl 0.1g、NaCl 0.5g、NaHCO30.5g、K2HPO4 0.2g、MgSO4·7H2O 0.1g、CaCl2Adding 0.1g of the mixture into 1000mL of distilled water, adjusting the pH value to 7 by using 10% of Na0H and 10% of HCI in percentage by mass, and uniformly stirring to obtain the photosynthetic bacteria liquid culture medium.
2. Enrichment culture of photosynthetic bacteria
200ml of photosynthetic bacteria liquid culture medium is prepared, the photosynthetic bacteria liquid culture medium is poured into a 250 ml conical flask for high-pressure steam sterilization, wherein the sterilization temperature is 115 ℃, the sterilization time is 30 minutes, and the conical flask is taken out and cooled to the room temperature. Transferring photosynthetic bacteria stored in the 7-ring solid culture medium into a conical flask by using an inoculating ring, uniformly mixing, wrapping the periphery of the conical flask by using gauze, placing the conical flask into an incubator, and performing illumination anaerobic culture for 48 hours under the condition that the light intensity is 2000lux, wherein the culture temperature is 30 ℃, so as to obtain the photosynthetic bacteria enrichment culture solution.
Preparation of photosynthetic bacteria immobilized filler
4. Preparation of coating solution
And adding polyvinylidene fluoride into the N, N-dimethylformamide solution, stirring and dissolving to obtain the polyvinylidene fluoride solution.
5. Adsorption of glass fiber balls to bacteria
Weighing 0.5g of glass fiber balls, washing, drying, and adding photosynthetic bacteria with the concentration of 2g/L for adsorption.
6. Preparation of immobilized fillers
Calculating the mass of the adsorbed bacteria to be 0.3g by a mass difference method, placing the glass fiber balls in the prepared polyvinylidene fluoride solution, stirring, shaking uniformly, taking out, placing in deionized water, standing for 4min, taking out, and washing with distilled water.
Fourthly, simulating high-salinity wastewater indexes: salinity of 5%, COD: 2900 mg/L.
Fifth, activity detection of immobilized filler
0.5g of immobilized filler is put into 200mL of high-salinity wastewater and treated for 72h under the conditions that the temperature is 30 ℃ and the illumination intensity is 2000 lux.
Two groups of experiments are set, and the difference is that the concentration of the high polymer in the coating solution is respectively 8% and 20%, and as can be seen from figure 2, the degradation of the high salinity wastewater COD by the immobilized photosynthetic bacteria with the concentration of the high polymer of 8% can reach 89%; the degradation of the high salinity wastewater COD by the immobilized photosynthetic bacteria with the high polymer concentration of 20 percent can reach 78 percent.
Comparative example 1:
cultivation of photosynthetic bacteria
1. Configuring photosynthetic bacteria liquid culture medium
Weighing yeast extract 0.5g, peptone 0.5g, and CH3COONa 3g、NH4Cl 0.1g、NaCl 0.5g、NaHCO30.5g、K2HPO4 0.2g、MgSO4·7H2O 0.1g、CaCl2Adding 0.1g of the mixture into 1000mL of distilled water, adjusting the pH value to 7 by using 10% of Na0H and 10% of HCI in percentage by mass, and uniformly stirring to obtain the photosynthetic bacteria liquid culture medium.
2. Enrichment culture of photosynthetic bacteria
200ml of photosynthetic bacteria liquid culture medium is prepared, the photosynthetic bacteria liquid culture medium is poured into a 250 ml conical flask for high-pressure steam sterilization, wherein the sterilization temperature is 115 ℃, the sterilization time is 30 minutes, and the conical flask is taken out and cooled to the room temperature. Transferring photosynthetic bacteria stored in the 7-ring solid culture medium into a conical flask by using an inoculating ring, uniformly mixing, wrapping the periphery of the conical flask by using gauze, placing the conical flask in an incubator, and performing illumination anaerobic culture for 48 hours under the condition that the light intensity is 2000lux, wherein the culture temperature is 30 ℃, so as to obtain the photosynthetic bacteria enrichment culture solution.
Secondly, simulating high-salinity wastewater indexes: salinity of 5%, COD: 2900 mg/L.
Thirdly, photosynthetic bacteria degrade high-salinity wastewater
0.3g of photosynthetic bacteria is put into 200mL of high-salinity wastewater and treated for 72 hours under the conditions that the temperature is 30 ℃ and the illumination intensity is 2000 lux.
As can be seen from FIG. 2, the suspended photosynthetic bacteria can degrade the COD of the high-salinity wastewater by 63%. Obviously lower than the degradation of the immobilized filler to COD.
Example 3:
firstly, preparing glass fiber balls: in accordance with example 1
Secondly, culturing photosynthetic bacteria: in accordance with example 1
Preparation of photosynthetic bacteria immobilized filler
1. Adding polysulfone (molecular weight: 65000Da) into N, N-dimethylformamide solution, stirring, and dissolving to obtain polysulfone solution; wherein the mass fraction of the polysulfone is 10%.
2. Adsorption of glass fiber balls to bacteria: in accordance with example 1
3. Preparation of immobilized fillers
Calculating the mass of the adsorbed bacteria to be 0.3g by a mass difference method, placing the glass fiber balls in the prepared polysulfone solution, stirring, shaking uniformly, taking out, placing in deionized water, standing for 3min, taking out, and washing with distilled water.
Fourthly, simulating indexes of the high-salinity wastewater: salinity of 7%, COD: 2900 mg/L.
Fourth, activity detection of immobilized filler
0.5g of immobilized filler is put into 200mL of high-salinity wastewater and treated for 72h under the conditions that the temperature is 30 ℃ and the illumination intensity is 2000 lux.
The degradation of the immobilized photosynthetic bacteria to the COD of the salt-containing wastewater can reach 91 percent.
Comparative example 2:
firstly, culturing photosynthetic bacteria: in accordance with example 3
Secondly, simulating high-salinity wastewater indexes: salinity of 7%, COD: 2900 mg/L.
Thirdly, photosynthetic bacteria degrade high-salinity wastewater
0.3g of photosynthetic bacteria is put into 200mL of high-salinity wastewater and treated for 72 hours under the conditions that the temperature is 30 ℃ and the illumination intensity is 2000 lux.
The degradation of suspended photosynthetic bacteria to the COD of the salt-containing wastewater is 75 percent.

Claims (5)

1. The composite photosynthetic bacteria immobilized filler is characterized by being prepared by the following method:
(1) culturing photosynthetic bacteria in a photosynthetic bacteria liquid culture medium under the condition of illumination and anaerobic conditions until the concentration is 1-2g/L in terms of dry cell mass to obtain a photosynthetic bacteria enrichment culture solution, adding glass fiber balls, and adsorbing the photosynthetic bacteria in pore channel structures of the glass fiber balls through the adsorption action of the glass fiber balls to obtain the photosynthetic bacteria adsorption glass fiber balls; the preparation method of the glass fiber ball comprises the following steps: sterilizing glass fibers, performing mechanical hot ligation on the glass fibers to prepare spheres with uniform sizes, and cleaning and drying the spheres to obtain the glass fiber spheres;
(2) adding the high polymer into an organic solvent, stirring and dissolving at the temperature of 20-40 ℃ to obtain a coating reagent; the mass fraction of the high polymer in the coating reagent is 8-20%; the organic solvent in the step (2) is N, N-dimethylformamide, N-dimethylacetamide or N-methylpyrrolidone; the high polymer is polysulfone or polyvinylidene fluoride;
(3) and (2) immersing the glass fiber balls adsorbing the photosynthetic bacteria obtained in the step (1) in the coating reagent prepared in the step (2), stirring, shaking uniformly, taking out, placing in deionized water, standing to form a film-shaped porous coating layer on the surfaces of the glass fiber balls, taking out, and washing with distilled water to obtain the composite photosynthetic bacteria immobilized filler.
2. The composite photosynthetic bacteria immobilized filler of claim 1, wherein: the final concentration of the photosynthetic bacteria liquid culture medium is as follows: 0.5g/L yeast extract and 0.5g/L, CH peptone3COONa 3 g/L、NH4Cl 0.1 g/L、NaCl 0.5 g/L、NaHCO30.5 g/L、K2HPO4 0.2 g/L、MgSO4·7H2O 0.1 g/L、CaCl20.1g/L, water as solvent, and initial pH of 7-9.
3. The composite photosynthetic bacteria immobilized filler of claim 1, wherein: the volume of the photosynthetic bacteria enrichment culture solution in the step (1) is 100-1000 mL/g based on the mass of the added glass fiber balls.
4. The composite photosynthetic bacteria immobilized filler of claim 1, wherein: the photosynthetic bacteria are Rhodopseudomonas (Rhodopseudomonas palustris) photosynthetic bacteria.
5. The use of the composite photosynthetic bacteria immobilized filler of claim 1 in degrading high-salinity wastewater COD.
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